Types | DnaRegion
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Roles | engineered_region
Composite
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Sequences | BBa_K1650007_sequence (Version 1)
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Description
The composite part contains a lac-inducible CDI operon which gives your strain the capability to inhibit growth of bacterial cells that express BamA in their outer membrane (most gram negative bacteria). Additionally the part contains mRFP1 (BBa_E1010) driven by another lac-inducible promoter as induction control. Moreover the composite part contains LacI.
IPTG-induced cells harboring BBa_K1650007 express CdiB which forms a beta-barrel structure. The CdiA protein is transported through CdiB and is displayed on the cell surface. When getting in contact with BamA of a target cell, the C-terminal domain is delivered into the target cell acting as pore forming protein thus destroying the proton motoric force of the target cell.
BBa_K1650007 could be used to improve transformation efficiency. Induced CDI+ cells would kill false positive cells.
Notes
BBa_K1650007 was toxic for cells in pSB1C3 backbone, presumably because of the high metabolic burden. In accordance with iGEM Headquarter, we submitted BBa_K1650007 in the low copy pSB4C5 backbone
Source
The CDI-operon was PCR amplified from genomic DNA of E.coli strain EC93. Other parts were taken from the registry.